Vps8 overexpression inhibits HOPS-dependent trafficking routes by outcompeting Vps41/Lt

  1. Péter Lőrincz  Is a corresponding author
  2. Lili Anna Kenéz
  3. Sarolta Tóth
  4. Viktória Kiss
  5. Ágnes Varga
  6. Tamás Csizmadia
  7. Zsófia Simon-Vecsei
  8. Gábor Juhász  Is a corresponding author
  1. Eötvös Loránd University, Hungary
  2. Hungarian Academy of Sciences, Hungary

Abstract

Two related multisubunit tethering complexes promote endolysosomal trafficking in all eukaryotes: Rab5-binding CORVET that was suggested to transform into Rab7-binding HOPS. We have previously identified miniCORVET, containing Drosophila Vps8 and three shared core proteins, which is required for endosome maturation upstream of HOPS in highly endocytic cells (Lorincz et al., 2016a). Here we show that Vps8 overexpression inhibits HOPS-dependent trafficking routes including late endosome maturation, autophagosome-lysosome fusion, crinophagy and lysosome-related organelle formation. Mechanistically, Vps8 overexpression abolishes the late endosomal localization of HOPS-specific Vps41/Lt and prevents HOPS assembly. Proper ratio of Vps8 to Vps41 is thus critical because Vps8 negatively regulates HOPS by outcompeting Vps41. Endosomal recruitment of miniCORVET- or HOPS-specific subunits requires proper complex assembly, and Vps8/miniCORVET is dispensable for autophagy, crinophagy and lysosomal biogenesis. These data together indicate the recruitment of these complexes to target membranes independent of each other in Drosophila, rather than their transformation during vesicle maturation.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files.

Article and author information

Author details

  1. Péter Lőrincz

    Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary
    For correspondence
    concrete05@gmail.com
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7374-667X
  2. Lili Anna Kenéz

    Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary
    Competing interests
    The authors declare that no competing interests exist.
  3. Sarolta Tóth

    Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary
    Competing interests
    The authors declare that no competing interests exist.
  4. Viktória Kiss

    Institute of Genetics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary
    Competing interests
    The authors declare that no competing interests exist.
  5. Ágnes Varga

    Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary
    Competing interests
    The authors declare that no competing interests exist.
  6. Tamás Csizmadia

    Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary
    Competing interests
    The authors declare that no competing interests exist.
  7. Zsófia Simon-Vecsei

    Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7909-4895
  8. Gábor Juhász

    Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary
    For correspondence
    szmrt@elte.hu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8548-8874

Funding

Magyar Tudományos Akadémia (LP-2014/2)

  • Gábor Juhász

Ministry of Human Capacities of Hungary (ÚNKP-18-4-ELTE-409)

  • Zsófia Simon-Vecsei

Magyar Tudományos Akadémia (PPD-222/2018)

  • Péter Lőrincz

Magyar Tudományos Akadémia (BO/00652/17)

  • Zsófia Simon-Vecsei

National Research, Development and Innovation Office of Hungary (GINOP-2.3.2-15-2016-00006)

  • Gábor Juhász

National Research, Development and Innovation Office of Hungary (GINOP-2.3.2-15-2016-00032)

  • Gábor Juhász

National Research, Development and Innovation Office of Hungary (K119842)

  • Gábor Juhász

National Research, Development and Innovation Office of Hungary (KKP129797)

  • Gábor Juhász

National Research, Development and Innovation Office of Hungary (PD124594)

  • Zsófia Simon-Vecsei

Ministry of Human Capacities of Hungary (ÚNKP-18-2-II-ELTE-32)

  • Lili Anna Kenéz

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Alexey J Merz, University of Washington School of Medicine, United States

Publication history

  1. Received: January 31, 2019
  2. Accepted: June 13, 2019
  3. Accepted Manuscript published: June 13, 2019 (version 1)
  4. Version of Record published: June 25, 2019 (version 2)

Copyright

© 2019, Lőrincz et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 1,927
    Page views
  • 329
    Downloads
  • 13
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Péter Lőrincz
  2. Lili Anna Kenéz
  3. Sarolta Tóth
  4. Viktória Kiss
  5. Ágnes Varga
  6. Tamás Csizmadia
  7. Zsófia Simon-Vecsei
  8. Gábor Juhász
(2019)
Vps8 overexpression inhibits HOPS-dependent trafficking routes by outcompeting Vps41/Lt
eLife 8:e45631.
https://doi.org/10.7554/eLife.45631

Further reading

    1. Cell Biology
    Agustin Leonardo Lujan, Ombretta Foresti ... Vivek Malhotra
    Research Article Updated

    We show that TANGO2 in mammalian cells localizes predominantly to mitochondria and partially at mitochondria sites juxtaposed to lipid droplets (LDs) and the endoplasmic reticulum. HepG2 cells and fibroblasts of patients lacking TANGO2 exhibit enlarged LDs. Quantitative lipidomics revealed a marked increase in lysophosphatidic acid (LPA) and a concomitant decrease in its biosynthetic precursor phosphatidic acid (PA). These changes were exacerbated in nutrient-starved cells. Based on our data, we suggest that TANGO2 function is linked to acyl-CoA metabolism, which is necessary for the acylation of LPA to generate PA. The defect in acyl-CoA availability impacts the metabolism of many other fatty acids, generates high levels of reactive oxygen species, and promotes lipid peroxidation. We suggest that the increased size of LDs is a combination of enrichment in peroxidized lipids and a defect in their catabolism. Our findings help explain the physiological consequence of mutations in TANGO2 that induce acute metabolic crises, including rhabdomyolysis, cardiomyopathy, and cardiac arrhythmias, often leading to fatality upon starvation and stress.

    1. Cell Biology
    2. Cancer Biology
    Chelsea U Kidwell, Joseph R Casalini ... Minna Roh-Johnson
    Research Article Updated

    Recent studies reveal that lateral mitochondrial transfer, the movement of mitochondria from one cell to another, can affect cellular and tissue homeostasis. Most of what we know about mitochondrial transfer stems from bulk cell studies and have led to the paradigm that functional transferred mitochondria restore bioenergetics and revitalize cellular functions to recipient cells with damaged or non-functional mitochondrial networks. However, we show that mitochondrial transfer also occurs between cells with functioning endogenous mitochondrial networks, but the mechanisms underlying how transferred mitochondria can promote such sustained behavioral reprogramming remain unclear. We report that unexpectedly, transferred macrophage mitochondria are dysfunctional and accumulate reactive oxygen species in recipient cancer cells. We further discovered that reactive oxygen species accumulation activates ERK signaling, promoting cancer cell proliferation. Pro-tumorigenic macrophages exhibit fragmented mitochondrial networks, leading to higher rates of mitochondrial transfer to cancer cells. Finally, we observe that macrophage mitochondrial transfer promotes tumor cell proliferation in vivo. Collectively these results indicate that transferred macrophage mitochondria activate downstream signaling pathways in a ROS-dependent manner in cancer cells, and provide a model of how sustained behavioral reprogramming can be mediated by a relatively small amount of transferred mitochondria in vitro and in vivo.